Screening for Biofuel Producing Clostridium Strains

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The use of fossil fuels for energy is convenient and affordable; however, there is always a demand of renewable energy due to the limited supplies of fuels. Fermentative Clostridium is a bacterium that is known to produce biofuel through ABE (acetone, butanol, and ethanol) fermentation. Butanol is one of the best potential candidates to replace gasoline for their similar carbon chain composition. Among Clostridium, C. beijerinckii was used prior to 1950s for butanol production. Our previous study has isolated 92 C. beijerinckii from horse fecal samples and environmental samples collected from three horse farms in southern California: Cal Poly Pomona, Mt. SAC, and a private farm in Glendora. To screen for strains that posses the ABE fermentation, three different sets of PCR primers were customized to detect the presence of genes encoding for alcohol dehydrogenase, acetate kinase, and butanol dehydrogenase, which are the enzymes involved in the production of alcohol, acetate, and butanol, respectively. PCR screening revealed that, of the 92 C. beijerinckii isolated that were screened, a total of 6 isolates were found to be positive for the three genes tested. Future studies will include the analysis of gene expression within the ABE fermentation pathway and its relationship to butanol production levels.